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目的 :观察用生物素标记的发夹寡核苷酸探针 (HPP)检测未成熟大鼠脑缺氧缺血后DNA双链断裂(DSB)的短暂空间改变 ,并与TdT介导的dUTP缺口末端标记 (TUNEL)的DNADSB比较。方法 :使 7d龄幼鼠缺氧缺血 ,用HPP位点标记显示DNADSB短暂空间形式 ,用微管相关蛋白 (MAP 2 )双标记评价HPP标记和脑损伤的关系 ,用双标记的TUNEL评价HPP标记的灵敏度。结果 :HPP标记局限在同侧半球MAP 2染色阴性区域 ;阳性细胞数与再灌注时间有关 ,在缺氧缺血后 2 4h ,阳性细胞数上升达到高峰 ,但在松果体细胞核变化更早 ,而海马CA3 区域细胞变化延迟。HPP和TUNEL双标记显示在再灌注早期HPP标记多于TUNEL标记。结论 :HPP标记显示出缺氧缺血后不同时间点凋亡细胞的典型变化 ,是检测凋亡改变的灵敏标记。
OBJECTIVE: To observe the transient spatial changes of DNA double-strand break (DSB) in immature rat brain after hypoxia and ischemia with biotin-labeled hairpin oligonucleotide probe (HPP) and compare with TdT-mediated dUTP gap DNADSB comparison of TUNEL. Methods: Hypoxic-ischemic neonatal rats (7-day-old) were subjected to hypoxia-ischemia. HPP sites were used to display the short form of DNA-DSB. The relationship between HPP-labeled and brain injury was evaluated by MAP 2 double labeling. Marked sensitivity. Results: The HPP marker was confined to the negative region of MAP 2 staining in the ipsilateral hemisphere. The number of positive cells was related to the reperfusion time. The number of positive cells peaked at 24 h after hypoxia and ischemia, The change of hippocampal CA3 cells is delayed. HPP and TUNEL double labeling showed that there were more HPP markers than TUNEL markers in the early stage of reperfusion. Conclusion: HPP labeling shows the typical changes of apoptotic cells at different time points after hypoxia-ischemia, which is a sensitive marker for detecting apoptosis.